Biotransformation processes metabolic activation

These studies with aldrin and antipyrine are sufficient to document the in vivo oxidative metabolic capability of mussels. Limits of activity have not been established, but they will be explored by further varying dose and the duration of the test period. With these limits established, the influence of environmental stressors such as salinity, and dissolved and/or suspended particulate matter in water on biotransformation will be assessed. If biotransformation processes are affected by these conditions, their measurement may provide results which can be diagnostically used as indicators of environmental quality. 

Because of the possible effects of active and carrier-mediated processes and metabolic biotransformation, the issue of tissue viability is important for in vitro buccal mucosal experiments. The barrier nature of the buccal mucosa resides in the upper layers of the epithelium, where unlike in the stratum corneum, the cells contain a variety of functional organelles [119, 122, 125, 150], and so tissue viability may be an important component of the barrier function of the tissue. Various methods have been employed to assess the viability of excised buccal mucosa, including measurement of biochemical markers, microscopic methods, and linearity of transport data [42], While biochemical methods, including measurement of adenosine 5 -triphosphate (ATP) levels and utilization of glucose, provide information on the metabolic activity of the tissue, this does not necessarily relate to the barrier function of the tissue. In excised rabbit buccal mucosa, levels of ATP were measured and found to decline by 40% in 6 h, and this correlated well with transmission electron microscopic evaluation of the tissue (intact superficial cells) [32], In addition, the permeability of a model peptide was unaltered up to 6 h postmortem, but at 8 h, a significant change in permeability was observed [32], These investigators therefore claimed that excised rabbit buccal mucosa could be used for diffusion studies for 6 h. 

The enzymatic route which a drug or poison follows in its metabolism is very specific to the xenobiotic itself. Substances with the same type of structures need not go through the same pathway. And the "active" form of the drug or the "toxic" form of the poison may occur either at the beginning or during the course of transformation. Usually synthetic combinations are not active or toxic but many of their precursors are. The following diagram illustrates the possibilities of a biotransformation process. 

Humans are exposed continuously and unavoidably to a myriad of potentially toxic chemicals that are inherently lipophilic and, consequently, very difficult to excrete. To effect their elimination, the human body has developed appropriate enzyme systems that can transform metabolically these chemicals to hydrophilic, readily excretable, metabolites. This biotransformation process occurs in two distinct phases. Phase I and Phase II, and involves several enzyme systems, the most important being the cytochromes P450. The expression of these enzyme systems is regulated genetically but can be modulated also other factors, such as exposure to chemicals that can either increase or impair activity. Paradoxically, the same xenobiotic-metabolizing enzyme systems also can convert biologically inactive chemicals to highly reactive intermediates that interact with vital cellular macromolecules and elicit various forms of toxicity. Thus, xenobiotic metabolism does not always lead to deactivation but can result also in metabolic activation with deleterious consequences. 

Bioavailability" is a quantitative measure of the utilization of an element under specific conditions, and includes mechanisms such as absorption, transport to a site of metabolic/toxic activity, biotransformation to a metabolically active/toxic form, retention/accumulation, and excretion (McKenzie in Nriagu, 1984). Bioavailable species of a pollutants can be increased by small particle size or volatility, aque-ous/lipid solubility, complexation, and - for metals - by elements or complexes which mimic essential nutrients and thus are handled by specific active transport processes. Assessment of pollutant bioavailability in sediments is difficult due the compexicity of the system, and the following questions have been raised with respect to toxic trace metals (Luoma, 1983 Salomons Forstner, 1984 Campbell et al., 1988) ... 

Another important point concerning the chemistry of the haptens must be underlined. Often, the hapten can be directly chemically reactive. Sometimes, the hapten needs preliminary metabolic activation (biotransformation). The reactions involved, in our organisms governed by breathing cell oxygenation, are essentially oxidative. This need for prior hapten oxidation is particularly true for metals involved in allergenic process. We know that the nickel in the form of Ni (11) (or NiO, nickel monoxide, or green nickel)... 

PEST. This code ( 3) was developed within the framework of Rensselaer Polytechnic Institute s CLEAN (Comprehensive Lake Ecosystem Analyzer) model. It includes highly elaborated algorithms for biological phenomena, as described in this volume (44). For example, biotransformation is represented via second-order equations in bacterial population density (Equation 5) in the other codes described in this section PEST adds to this effects of pH and dissolved oxygen on bacterial activity, plus equations for metabolism in higher organisms. PEST allows for up to 16 compartments (plants, animals, etc.), but does not include any spatially resolved computations or transport processes other than volatilization. 

Identification of metabolic reactions at an early phase can significantly affect the drug discovery process, because bioavailability, activity, toxicity, distribution and final elimination all depend on metabolic biotransformations [1], Once obtained, this information can help researchers judge whether or not a potential candidate should be eliminated from the pipeline or modified to reduce the affinity for CYP antitarget enzymes. 

As mentioned above, bioavailability is the degree to which a drug reaches the intended site of action. The amount of drug that reaches systemic circulation will depend on the processes of absorption, distribution, and biotransformation (when the route of administration exposes the drug to first-pass metabolism). Pharmacokinetics are often linear and when they are nonlinear it is often due to a saturation of protein binding, metabolism, or active renal transport. 

Many reported biotransformations are initially only demonstrated on a very small scale, the substrates or products may be subject to competing reactions if other enzymes are present (this can be a serious issue in whole-cell biocatalysis), or the desired enzyme is insufficiently active or produced in low levels. For many biotransformations a little care and attention is needed in the growth of the microbe to achieve the desired results. Production of a specific enzyme from a microbe can often be increased by growing the cells in the presence of a very small concentration (typically micromolar) of an inducer. The inducer could be a natural enzyme substrate, a substrate mimic or a molecule which is in some way associated with a substrate s availability or role in metabolism. This process is called induction and represents a genetic switch which cells use to respond... 

The introduction of fluorine atoms in a molecule can be used to modify the processes and the rates of metabolism of the drug, in order to extend the plasma half-life or avoid the formation of toxic metabolites. Due to the properties of the fluorine atom, in particular its electronic effects, it may interact differently during the biotransformation steps, according to the type of processes involved (oxidative, reductive, hydrolytic, etc), which allow the clearance of the exogen molecule (i.e., the elimination of the active substance from the organism). 

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